在无序结合基因中,连接器的长度和组成调节了多价值蛋白相互作用开关的激烈性和可逆性
Kiran Sankar Chatterjee1, Maria A Martinez-Yamout1, H Jane Dyson1
1Department of Integrative Structural and Computational Biology and Skaggs Institute of Chemical Biology, Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037.
本质上有障碍的蛋白质调节细胞对低氧 (低氧) 的反应. 这项研究揭示了缺氧诱导因子1-alpha (HIF-1α) 的突变如何微调其与协同激活剂的相互作用,影响缺氧开关.
科学领域:
- 分子生物学分子生物学
- 生物化学 生化学
- 细胞生物学 细胞生物学
背景情况:
- 本质上有障碍的蛋白质在缺氧期间控制基因表达.
- CITED2作为反抑制剂,通过将HIF-1α从CBP/p300联合激活器中取代来终止低氧反应.
- 这种开关的单向性取决于变化和热力学合.
研究的目的:
- 调查约束性合作在HIF-1α/TAZ1相互作用中的作用.
- 了解链接器突变如何影响HIF-1α的TAZ1结合基因之间的热力学合.
- 为了阐明低氧开关精确调节的分子基础.
主要方法:
- 在HIF-1α激活域链接序列的突变发生.
- 原生凝电泳以评估HIF-1α/TAZ1结合亲和力.
- 光异构性竞争测定和核磁共振 (NMR) 谱法来测量CITED2竞争.
主要成果:
- 特定的链接器突变增强了HIF-1α对TAZ1域的亲和力.
- 突变的HIF-1α变种比野生类型更有效地与CITED2竞争TAZ1结合.
- 连接器的灵活性 (甘氨酸存在) 和静电相互作用 (氨酸侧链) 对TAZ1亲和力和CITED2竞争至关重要.
结论:
- 该研究提供了对低氧转录反应调节的分子见解.
- 在HIF-1α中链接序列的修改可以调节其与协同激活剂的相互作用,影响低氧开关.
- 在HIF-1α链接器中的灵活性和静电相互作用是TAZ1结合和CITED2的竞争性取代的关键决定因素.
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